Funding is requested for an automated, high throughput (HT) microscope manufactured by Q3DM/Beckman Coulter. The instrument (known as IC-100, previously Eidaq 100) automatically scans and acquires thousands of brightfield or fluorescent images of cells in multiwell dishes. Sophisticated software analyzes the images and calculates many measurements of the intensity and localization of fluorescent or brighfield objects within individual cells. Thus, it is possible to evaluate a range of subcellular changes, such as gene or protein expression, protein or organelle trafficking, and signal transduction, in response to many reagents or culture conditions on living or fixed cells. The sole accessory requested is a robotic arm and incubator to permit the automatic loading of multiwell cell culture plates onto the microscope. This will expand the capacity of the instrument by enabling unattended operation. The user base is large and diverse, representing the Burnham Institute, Salk Institute and University of California, San Diego and the projects support grants from NAI, NCI, NHLBI, NIDDK, NIAID, NIMH and NIGMS. Seven projects from major users are highlighted in section B of this proposal and represent disease-oriented research in cardiology, diabetes, cancer, and infectious disease. The HT microscope will be managed as part of a shared facility for cell analysis that also includes a high speed cell sorter (Beckman FACSVantage SE) and analytical cytometers. This facility and HT microscopy is a key part of a larger initiative to screen libraries of small molecules, proteins and genes for applications in cancer, infectious disease and regenerative medicine. The Institutional commitment to this initiative is substantial and is reflected in the purchase of advanced robotics and chemical libraries and the hiring of technical staff and faculty (letters from the Institute's CFO and Cancer Center Director expressing support for the initiative are supplied in the appendix). [unreadable] [unreadable]